日本七鳃鳗口腔腺分泌蛋白BGSP-2的序列进化分析、基因克隆及诱导表达

Phylogenetic analysis and purification of Lampetra japonica oral gland secreted BGSP-2 protein

从实验构建的七鳃鳗口腔腺cDNA文库中获取BGSP-2(buccal gland secretion protein)基因片段,并对其进行生物信息学分析.RT-PCR扩增,已获得日本七鳃鳗口腔腺BGSP-2蛋白的全序列cDNA,将目的基因与pET23b载体连接,进一步转化至Rosetta表达菌株中,筛选阳性克隆,对包涵体进行变性、复性与纯化.结果显示已成功获得成分均一的活性蛋白.

The genome segment of BGSP-2 was taken from the buccal gland of Lampetra japonica according to the information from cDNA library and the primary analysis of expressed sequence tags（EST） which constructed in our lab,and analyzed by bioinformations.The whole cDNA of BGSP-2 was obtained by RT-PCR.The BGSP-2 was transformed into the vector of pET23b,furthermore,converted into strains of Rosetta and selected positive clones.To obtain the soluble homogeneous protein,Denaturation,renaturation and purification of the inclusionbodies from recombinant BGSP-2 was carried out.